What question did this study set out to answer?

To purify fatty acid cyclooxygenase from sheep vesicular glands and determine its molecular properties and iron cofactor requirements.

September 1, 1976Journal of Biological ChemistryOpen Access

Purification of the cyclooxygenase that forms prostaglandins. Demonstration of two forms of iron in the holoenzyme.

View Full Paper

Ask AI

Bookmark

To purify fatty acid cyclooxygenase from sheep vesicular glands and determine its molecular properties and iron cofactor requirements.
Purified cyclooxygenase from sheep vesicular glands using detergent solubilization, ammonium sulfate fractionation, DEAE-cellulose chromatography, flurbiprofen-Sepharose affinity chromatography, isoelectric focusing, and gel filtration.
Characterized protein purity, molecular weight, and aspirin binding using SDS-PAGE and radioactive [3H]acetylsalicylic acid labeling.
Evaluated catalytic activity and iron status using [1-14C]arachidonic acid conversion assays and aerobic optical absorption spectrophotometry with and without added hemin.
Purified the enzyme to a specific activity of 46,000 units/mg protein, yielding a single 70,000 molecular weight band on SDS-PAGE that selectively co-migrated with radioactive acetyl groups.
Catalyzed the complete double-oxygen insertion needed to generate prostaglandin PGG2 from arachidonic acid, converting into pure PGF2alpha in the presence of stannous chloride.
Identified significant nonheme iron along with a faint 412 nm heme absorption peak, showing that enzyme activation occurs upon binding exogenous hemin to form a 407 nm heme-protein complex.

Discussion

No takes yet. Share an insight, caveat, or question.

Implication

Biochemical study demonstrates isolation of prostaglandin-forming cyclooxygenase from sheep vesicular glands, revealing dual forms of iron essential for enzyme activation.

Cite This Study

Hemler et al. (1976) studied this question.

synapsesocial.com/papers/6a1c11ef00ee29383e9d686d https://doi.org/10.1016/s0021-9258(17)33098-3

View Full Paper

Ask AI

Bookmark

Implication

Biochemical study demonstrates isolation of prostaglandin-forming cyclooxygenase from sheep vesicular glands, revealing dual forms of iron essential for enzyme activation.

Purification of the cyclooxygenase that forms prostaglandins. Demonstration of two forms of iron in the holoenzyme.

Key Points

Discussion

Implication

Cite This Study

Purification of the cyclooxygenase that forms prostaglandins. Demonstration of two forms of iron in the holoenzyme.

Key Points

Discussion

Implication

Cite This Study

Also Consider

Also Consider